Pressure-Dependent Elevation of Vasoactive Intestinal Peptide Level in Chicken Choroid.

PURPOSE: Autonomic control is important in maintaining ocular integrity. As recent data suggested that intrinsic choroidal neurons (ICN), an intrinsic choroidal autonomic control, may regulate choroidal thickening via release of the vasodilative vasoactive intestinal peptide (VIP), it was the aim of the study to investigate the level of choroidal VIP (VIPchor) in the presence of an increased atmospheric pressure in a chicken model. METHODS: Chicken choroidal whole mounts were exposed to ambient pressure (n = 20) and 40 mm Hg (n = 20) in a PC-controlled, open chamber system for 24 and 72 h, respectively. The VIP concentration was analyzed by ELISA, and the total protein concentration was measured by the BCA assay. Statistical analysis was done using an unpaired two-tailed t-test. RESULTS: The pressurization systems enabled choroidal whole mount pressurization (40 mm Hg) with humidifying, pressure, temperature, and gas exchange. Overall, the VIPchor level concentration was significantly increased at 40 mmHg compared to the ambient pressure (30.09 ± 7.18 pg vs. 20.69 ± 3.24 pg; p < 0.0001). Subgroup analysis yielded a significantly increased VIPchor level at 40 mmHg compared to the ambient pressure after 24 h (28.42 ± 6.03 pg vs. 20.76 ± 4.06 pg; p = 0.005) and 72 h (31.77 ± 7.82 pg vs. 20.61 ± 2.12 pg; p = 0.002), respectively. The VIPchor elevation at 40 mm Hg ranged between 1.37- (24 h) and 1.54-fold (72 h) compared to the ambient pressure. No difference was observed between the VIPchor level at 24 h and 72 h (p > 0.05). CONCLUSIONS: The increase of the total choroidal VIP level, representing the intracellular VIP content, in the presence of an increased ambient pressure argues for a retention of VIP within the neurons, decreasing both vasodilatation and, consequently, choroid thickness. This finding might be a passive or even active function of ICN in the regulation of choroidal thickness, ocular integrity and IOP.

Endophthalmitis: Epidemiology, Causing Agents, Therapy and Visual Outcome with Special Focus on Glaucoma Patients. / Endophthalmitis: Ursachen, Erreger, Therapie und Visusverlauf mit Fokus auf Glaukompatienten.

BACKGROUND: Endophthalmitis is one of the most serious emergencies in ophthalmology. In order to reduce its prevalence, it is important to have a proper understanding of potential risk. Surgical therapy with targeted, pathogen-specific medication and an intact immune system are fundamental for preserving visual acuity. As it is unclear whether an unfavourable course is more likely in the presence of underlying ocular disease, a comparison was made between glaucoma patients (G) and non-glaucoma patients (NG) in terms of causative factors, pathogens, treatment and visual acuity. Since a potential alteration of the local immune system in glaucoma has been described, it is of interest to determine whether the clinical course of endophthalmitis in glaucoma patients differ from that of non-glaucoma patients. PATIENTS AND METHODS: A retrospective analysis of 75 eyes (13 G, 62 NG) who underwent treatment and surgery following a diagnosis of endophthalmitis in the Department of Ophthalmology, University of Erlangen-Nuremberg has been evaluated over a period of 5 years. Clinical characteristics, surgical treatment, microbial spectrum and visual acuity in glaucoma and non-glaucoma eyes were investigated. RESULTS: Severe visual impairment (44%) with inflammation of the anterior chamber (62.7%), hypopyon (52%) and reduced (40%) or absent view (26.7%) of the fundus were predominantly present at first diagnosis in all patients. Previous eye surgery was observed in a total of 53%, primarily cataract surgery. Gram-positive cocci were seen as the most common causative agent in both groups, (G: 23.1%; NG: 38.7%), whereas other rare pathogens were present only in glaucomatous eyes. Pars plana vitrectomy was performed in 76% and enucleations in 20% of all patients, with the latter significantly more common in glaucomatous eyes (p = 0.01). A significant postoperative improvement in visual acuity was achieved in non-glaucoma patients (p < 0.001); visual acuity was worse in glaucomatous eyes. CONCLUSION: Although rare, early diagnosis and treatment of endophthalmitis is crucial in terms of prognosis. In the present cohort, worse visual acuity outcomes were obtained in glaucoma patients in comparison to non-glaucoma patients.

Immunohistochemical detection of urea transporter-A in the tear-producing part of the lacrimal system.

OBJECTIVE: Urea constitutes a physiological and presumably well-regulated constituent of tear fluid. Its lacrimal concentration is significantly decreased in dry eye disease. Urea homeostasis within the tear fluid may also depend on the expression of urea transporters. The present study reports on the expression patterns of urea transporter A (UT-A) in the cells and tissues of the ocular surface and the lacrimal glands. METHODS: UT-A immunohistochemistry was performed on 5 µm paraffin sections of paraformaldehyde-fixed human, porcine, and murine corneas, eyelids, and lacrimal glands (n = 5 each). RESULTS: UT-A immunostaining was largely comparable in all three species. UT-A signals were detected in the corneal epithelium and endothelium, in the conjunctival epithelium, in the acinar cells and excretory ducts of the lacrimal gland, Meibomian gland, and in the glands of Moll and Zeis. The Meibomian glands and the glands of Zeis exhibited a marked UT-A-positive staining in the basal cells of the alveolar epithelia and in the ductal epithelia. CONCLUSION: UT-A shows comparable expression patterns to UT-B (previous study) at the ocular surface and in the lacrimal glands, as determined by immunohistochemistry. The presence of both urea transporters in the lacrimal functional unit suggests that they are essential for the normal function of the lacrimal system and the integrity of the tear film. Potential alterations in urea transporter expression might be associated with the significant reduction of urea found in the tear fluid of dry eye patients. They may thus play an important role in the pathogenesis of dry eye disease.

Urea transporter-B expression on the ocular surface and in the lacrimal glands.

OBJECTIVE: Urea is a component of tear fluid showing a significantly decreased concentration in dry eye disease. The urea content of tear fluid may depend on urea transporters. The purpose of this study was to examine the expression of urea transporter B (UT-B) at the ocular surface and in the lacrimal glands. METHODS: UT-B protein and mRNA expression was investigated in human, porcine, and murine samples. Immunohistochemical staining for UT-B was performed on paraffin sections of human, porcine, and murine corneas, eyelids, and lacrimal glands (n = 5 each). Reverse transcriptase polymerase chain reaction was conducted to detect UT-B mRNA in human and murine cornea, conjunctiva, Meibomian gland, and lacrimal gland (n = 5 each). RESULTS: UT-B protein expression was comparable in all three species. It was found in the corneal epithelium and endothelium, in the conjunctival epithelium, in the end pieces and excretory ducts of the lacrimal gland, Meibomian gland, and in the glands of Moll and Zeis. The glands of Zeis and the Meibomian glands showed intense UT-B signals in the basal layers of the alveolar epithelia and in the cells of the ductal epithelia. UT-B mRNA was detected in all samples analyzed. CONCLUSION: UT-B is expressed by the cells and tissues of the ocular surface and in the lacrimal glands. Potential changes in urea transporter expression might have implications for the pathogenesis of dry eye disease. Since comparable results were obtained for all species investigated, the presented findings may open the door for DED-relevant experimentation on porcine and murine model systems.

Case Report: Neutralization of Autoantibodies Targeting G-Protein-Coupled Receptors Improves Capillary Impairment and Fatigue Symptoms After COVID-19 Infection.

Clinical features of Coronavirus disease 2019 (COVID-19) are caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Acute infection management is a substantial healthcare issue, and the development of long-Covid syndrome (LCS) is extremely challenging for patients and physicians. It is associated with a variety of characteristics as impaired capillary microcirculation, chronic fatigue syndrome (CFS), proinflammatory cytokines, and functional autoantibodies targeting G-protein-coupled receptors (GPCR-AAbs). Here, we present a case report of successful healing of LCS with BC 007 (Berlin Cures, Berlin, Germany), a DNA aptamer drug with a high affinity to GPCR-AAbs that neutralizes these AAbs. A patient with a documented history of glaucoma, recovered from mild COVID-19, but still suffered from CFS, loss of taste, and impaired capillary microcirculation in the macula and peripapillary region. He was positively tested for various targeting GPCR-AAbs. Within 48 h after a single BC 007 treatment, GPCR-AAbs were functionally inactivated and remained inactive during the observation period of 4 weeks. This observation was accompanied by constant improvement of the fatigue symptoms of the patient, taste, and retinal capillary microcirculation. Therefore, the removal of GPCR-AAb might ameliorate the characteristics of the LCD, such as capillary impairment, loss of taste, and CFS.

Retinal Microcirculation as a Correlate of a Systemic Capillary Impairment After Severe Acute Respiratory Syndrome Coronavirus 2 Infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), affects the pulmonary systems via angiotensin-converting enzyme-2 (ACE-2) receptor, being an entry to systemic infection. As COVID-19 disease features ACE-2 deficiency, a link to microcirculation is proposed. Optical coherence tomography angiography (OCT-A) enables non-invasive analysis of retinal microvasculature. Thus, an impaired systemic microcirculation might be mapped on retinal capillary system. As recent OCT-A studies, analyzing microcirculation in two subdivided layers, yielded contrary results, an increased subdivision of retinal microvasculature might offer an even more fine analysis. The aim of the study was to investigate retinal microcirculation by OCT-A after COVID-19 infection in three subdivided layers (I). In addition, short-term retinal affections were monitored during COVID-19 disease (II). Considering (I), a prospective study (33 patientspost-COVID and 28 controls) was done. Macula and peripapillary vessel density (VD) were scanned with the Spectralis II. Macula VD was measured in three layers: superficial vascular plexus (SVP), intermediate capillary plexus (ICP), and deep capillary plexus (DCP). Analysis was done by the EA-Tool, including an Anatomical Positioning System and an analysis of peripapillary VD by implementing Bruch's membrane opening (BMO) landmarks. Overall, circular (c1, c2, and c3) and sectorial VD (s1-s12) was analyzed. Considering (II), in a retrospective study, 29 patients with severe complications of COVID-19 infection, hospitalized at the intensive care unit, were monitored for retinal findings at bedside during hospitalization. (I) Overall (p = 0.0133) and circular (c1, p = 0.00257; c2, p = 0.0067; and c3, p = 0.0345). VD of the ICP was significantly reduced between patientspost-COVID and controls, respectively. Overall (p = 0.0179) and circular (c1, p = 0.0189) peripapillary VD was significantly reduced between both groups. Subgroup analysis of hospitalized vs. non-hospitalized patientspost-COVID yielded a significantly reduced VD of adjacent layers (DCP and SVP) with increased severity of COVID-19 disease. Clinical severity parameters showed a negative correlation with VD (ICP) and peripapillary VD. (II) Funduscopy yielded retinal hemorrhages and cotton wool spots in 17% of patients during SARS-CoV-2 infection. As VD of the ICP and peripapillary regions was significantly reduced after COVID-19 disease and showed a link to clinical severity markers, we assume that the severity of capillary impairment after COVID-19 infection is mapped on retinal microcirculation, visualized by non-invasive OCT-A.

Mouth-nose masks impair the visual field of healthy eyes.

BACKGROUND: Mouth-nose masks have been requested to prevent the transmission of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The aim of the present study was to investigate, if wearing a mouth-nose mask impairs the visual field function in normals. METHODS: Thirty eyes of 30 subjects were recruited for the present study. White-on-white perimetry (OCTOPUS 900; 90°) was done and sensitivity was analysed in 14 defined test points (P1-P14, inferior visual field) under 3 different test conditions while the subjects were wearing a mouth-nose mask: (I) 1.5 cm under the lower eyelid, nose clip not used (position1.5cm_no_clip); (II) 1.5 cm under the lower eyelid, nose clip correctly positioned (position1.5cm_with_clip); (III) 0.5 cm under the lower eyelid, nose clip correctly positioned (position0.5cm_with_clip). All data were compared to sensitivity without wearing a mouth-nose mask (reference). Mean Δ was calculated, being the difference between the results of each test condition and reference, respectively. RESULTS: Sensitivity was significantly different between position1.5cm_no_clip and reference at 10 test points (p<0.05). Sensitivity at test point P7 was significantly different between position1.5cm_with_clip and position0.5cm_with_clip compared to reference (p<0.001), respectively. Mean Δ increased while wearing a mask at P7: position1.5cm_with_clip (-8.3 dB ± 7.3 dB) < position0.5cm_with_clip (-11.3 dB ± 9.5 dB) < position1.5cm_no_clip (-20.1 dB ± 7.6 dB). CONCLUSION: Visual field function was observed to be significantly impaired in the inferior-nasal sector while persons were wearing a mouth-nose mask, especially when the nose clip was not correctly used.

Infrared thermography: different options of thermal eyelid warming.

PURPOSE: Current therapies of Meibomian gland dysfunction (MGD) include thermal eyelid warming. It was the aim of the study to investigate the temperature run after eyelid warming using 3 different techniques (hot compresses, Blephasteam® (Thea Pharma GmbH, France), and having a sauna) in patients with MGD compared with normal subjects by infrared thermography. METHODS: A prospective case-control study was done. Temperature profiles of the eyelids were investigated in 93 subjects (49 normals, 44 MGD patients) after warming of the eyelids by 3 methods: hot compresses, Blephasteam®, and having a sauna. Temperature runs of the eyelids were measured with an infrared thermal imaging camera (VarioCAM® HD research 675/30 mm, InfraTec GmbH) at baseline and after eyelid warming for 10 min. Statistical analysis were done by Wilcoxon test or t tests for unpaired samples. RESULTS: The initial eyelid temperature was significantly increased after the use of Blephasteam® compared with hot compresses in MGD and normal patients (p < 0.001). Having a sauna showed a similar warming effect of the eyelids than Blephasteam® in normals and MGD patients (p > 0.05). Additionally, the warming effect of the eyelids after having a sauna was significantly longer than after the use of Blephasteam® in MGD (p = 0.016) and normal patients (p = 0.01). CONCLUSION: Eyelid temperature after having a sauna was similar to commonly used warming devices; yet, the duration of the eyelid warming effect was longer. Thus, having a sauna might be an alternative option for warming of the eyelids.

ICare Pro: Age Dependent Effect of Central Corneal Thickness on Intraocular Pressure in Glaucoma and Ocular Hypertension Patients.

Purpose: Measurement of the exact intraocular pressure (IOP) is essential in glaucoma diagnosis and follow-up, thus all therapeutic options affect IOP in order to win sighted lifetime. As it is known that corneal properties of glaucoma patients differ from normal subjects, the present study aimed to investigate the influence of CCT on rebound tonometry (ICT, ICare Pro) in glaucoma and ocular hypertension patients in dependency of age additionally considering different times of day.Methods: Three hundred sixty-two eyes of 190 subjects were included: 339 open-angle glaucoma and 23 ocular hypertension. IOP was measured at 5 different times of day (6 a.m., 12 a.m., 4 p.m., 9 p.m., and 0 p.m.) by Goldmann applanation tonometry (GAT) and Icare Pro rebound tonometry in a sitting position. Central corneal thickness was measured by central ultrasonic pachymetry (Pachymeter SP-100). Δ ICT was calculated as the difference of GAT, corrected according to age and CCT, and ICT, respectively at each time point.Results: All different GAT time points data correlated significantly (p < .05) with ICT time points. An age effect was observed on overall ICT (p = .02). A decrease of ICT was observed with increasing age. The within differences among ICT repeated measurements were significant as well. Additionally, repeated means of Δ ICT correlated significantly with age and CCT. Intercepts and coefficients were offered for each time point, respectively. GLM model yielded a relation between MD (dependent variable) and age together with CCT (age: p < .0001) and (CCT: p = .043).Conclusions: IOP measurements with ICare Pro were shown to be dependent on age, CCT and time of day in glaucoma and ocular hypertension patients. Thus, aging, corneal biomechanical properties and circadian rhythms should be taken into consideration when adjusting IOP.

[MIGS-off-label option for treatment-refractory steroid-induced ocular hypertension]. / MIGS ­ eine Off-label-Option für eine therapierefraktäre, steroidinduzierte okuläre Hypertension.

Steroid-induced ocular hypertension (SIOH), which cannot be regulated by topical antiglaucoma agents, is a rare complication after intravitreal implantation of diverse cortisone derivatives. This case report presents a therapeutic approach for treatment-refractory SIOH after intravitreal implantation of fluocinolone acetonide using various minimally invasive glaucoma surgery (MIGS) techniques.

Mutations in PIK3C2A cause syndromic short stature, skeletal abnormalities, and cataracts associated with ciliary dysfunction.

PIK3C2A is a class II member of the phosphoinositide 3-kinase (PI3K) family that catalyzes the phosphorylation of phosphatidylinositol (PI) into PI(3)P and the phosphorylation of PI(4)P into PI(3,4)P2. At the cellular level, PIK3C2A is critical for the formation of cilia and for receptor mediated endocytosis, among other biological functions. We identified homozygous loss-of-function mutations in PIK3C2A in children from three independent consanguineous families with short stature, coarse facial features, cataracts with secondary glaucoma, multiple skeletal abnormalities, neurological manifestations, among other findings. Cellular studies of patient-derived fibroblasts found that they lacked PIK3C2A protein, had impaired cilia formation and function, and demonstrated reduced proliferative capacity. Collectively, the genetic and molecular data implicate mutations in PIK3C2A in a new Mendelian disorder of PI metabolism, thereby shedding light on the critical role of a class II PI3K in growth, vision, skeletal formation and neurological development. In particular, the considerable phenotypic overlap, yet distinct features, between this syndrome and Lowe's syndrome, which is caused by mutations in the PI-5-phosphatase OCRL, highlight the key role of PI metabolizing enzymes in specific developmental processes and demonstrate the unique non-redundant functions of each enzyme. This discovery expands what is known about disorders of PI metabolism and helps unravel the role of PIK3C2A and class II PI3Ks in health and disease.

VIP changes during daytime in chicken intrinsic choroidal neurons.

PURPOSE: Ocular autonomic control is mediated by sympathetic and parasympathetic nerve fibres. Their interactions are complemented by primary afferent nerve fibers of and intrinsic choroidal neurons (ICN). As the vasodilatative neuropeptide, vasoactive intestinal peptide (VIP), is expressed in extrinsic and intrinsic ocular neurons, it is of special interest in ophthalmic research. Since circadian changes of ocular blood flow are known in humans and birds, this study aimed at investigating VIP expression at different daytimes in chicken choroid, the preferred model species in ICN research. METHODS: 12 eyes of 12 chickens were retrieved, slaughtered at 8.00-9.30 a.m. (n = 6) and 8.00 p.m. (n = 6), respectively, and choroidal wholemounts were prepared for immunofluorescence of VIP. VIP-positive ICN of both groups were quantified and density of VIP-positive axons assessed semi-quantitatively. In 28 additional eyes retrieved in the morning (n = 14) and evening (n = 14), choroidal VIP content was determined by ELISA. Morning and evening data were analyzed statistically. NADPH-diaphorase (NADPH-d, ICN cell marker) was done at additional 12 whole mount choroids of 12 chicken, retrieved in the morning (n = 6) and evening (n = 6). RESULTS: (1) Numbers of VIP positive neurons differed significantly between morning: (239.17 ±â€¯113.9) and evening: (550.83 ±â€¯245.7; p = 0.018). (2) Numbers of VIP-positive perikarya were significantly more accumulated in the temporal part of the choroid in the evening than in the morning (p = 0.026). (3) VIP positive axon density was found to be similar throughout the choroid in the morning and evening. (4) Number of NADPH-d positive neurons was not significantly different between morning (848.8 ±â€¯399.5) and evening (945.8 ±â€¯622.1, p > 0.05). (5) ELISA demonstrated a significant difference of VIP content (p = 0.012) in tissues harvested in the morning (145.41 ±â€¯43.3 pg/ml) compared to evening (221.44 ±â€¯106.3 pg/ml). CONCLUSIONS: As VIP positive axon density was similar in the morning and the evening throughout the choroid, PPG and ICN seemed to contribute equally to the axon network. Yet, changes in the total choroidal VIP content, the numbers of VIP positive perikarya, reflecting the intracellular VIP content, and their topographical distribution at two different days-times argue for a different status of activation of both neuronal sources in contrast to the equal amount of NADPHD-d positive neurons. The higher VIP content in the evening, compared to the morning, correlates with a known circadian rhythm of a lower IOP and a higher choroidal thickness at night. Thus, these changes may argue for a potential role of ICN in the regulation of ocular homeostasis and integrity.

Follicular Conjunctivitis due to Chlamydia felis-Case Report, Review of the Literature and Improved Molecular Diagnostics.

A 29-year-old woman presented with unilateral, chronic follicular conjunctivitis since 6 weeks. While the conjunctival swab taken from the patient's eye was negative in a Chlamydia (C.) trachomatis-specific PCR, C. felis was identified as etiological agent using a pan-Chlamydia TaqMan-PCR followed by sequence analysis. A pet kitten of the patient was found to be the source of infection, as its conjunctival and pharyngeal swabs were also positive for C. felis. The patient was successfully treated with systemic doxycycline. This report, which presents one of the few documented cases of human C. felis infection, illustrates that standard PCR tests are designed to detect the most frequently seen species of a bacterial genus but might fail to be reactive with less common species. We developed a modified pan-Chlamydia/C. felis duplex TaqMan-PCR assay that detects C. felis without the need of subsequent sequencing. The role of chlamydiae-specific serum antibody titers for the diagnosis of follicular conjunctivitis is discussed.

Presence of HLA-B27 is associated with changes of serum levels of mediators of the Wnt and hedgehog pathway.

BACKGROUND: HLA-B27 is present in 5% of the Caucasian population and is strongly associated with the development of spondyloarthritis (SpA), a disease characterized by inflammation and substantial bone changes. We hypothesized that the presence of HLA-B27 in itself is associated with alterations of key regulatory of bone homeostasis. METHODS: Sera of 241 individuals were assessed for the serum levels of Wnt pathway regulators, sclerostin and dickkopf (Dkk)-1 as well as Indian hedgehog (IHH) and collagen type I cleavage products (CTX1). Of the 151 HLA-B27+ subjects, 31 had SpA, 30 had anterior uveitis, 30 were healthy individuals and 60 healthy siblings of patients with SpA. RESULTS: Sclerostin levels were significantly (P<0.001) lower in HLA-B27+ subjects (314±21pg/mL) compared to HLA-B27 negative controls (mean±SEM: 492±30pg/mL), no matter if subjects were either healthy, or affected by SpA or uveitis. Similar results were found for Dkk-1. No differences between the groups with respect to the bone resorption marker CTX1 were found. In contrast, IHH levels were significantly (P<0.001) higher in the carriers of HLA-B27 than in the negative controls. CONCLUSIONS: Changes in key regulators of the Wnt pathway as well as IHH, a molecule regulating endochondral ossification, are found in HLA-B27 carriers, independent if they were healthy or affected by uveitis or SpA.

Retinopathy in a patient with acute Epstein-Barr virus infection: follow-up analysis using spectral domain optical coherence tomography.

PURPOSE: To describe the clinical findings, diagnostics, and differential diagnosis in a patient with retinopathy in acute systemic Epstein-Barr virus (EBV) infection. METHODS: Description of the clinical course of the EBV retinopathy was based on the medical record, photographs, and visual fields of the patient. Retinal morphology was visualized using spectral domain optical coherence tomography (Heidelberg Engineering), and fluorescein angiography. Multiple serologic tests were performed to exclude different infectious agents. A 30-year-old male patient presented with a focal scotoma in his right visual field. One week ago, he suffered from high fever, joint pain, neck stiffness, and hepatitis; 2 weeks later, he had returned from a trip to Malaysia. RESULTS: Visual acuity was 20/20 in both eyes. Fundoscopy showed a paravascular sharp-edged white lesion with a corresponding crescent-shaped retinal nerve fiber defect. Serology revealed high titers for EBV immunoglobulin M (IgM), but no EBV immunoglobulin G (IgG), indicating an acute infectious mononucleosis. Other reasons for noninfectious or infectious retinochoroiditis including tropical microorganisms could be excluded serologically and by imaging. Treatment with systemic acyclovir and prednisolone acetate eye drops was initiated leading to recovery of systemic and ocular findings. CONCLUSION: Acute systemic EBV infection can affect the retina leading to focal thickening of the inner retinal layers because of focal microinfarction (cotton-wool spot) or focal infectious infiltration. Spectral domain optical coherence tomography is capable of detecting changes in retinal morphology in such cases.

Choroidal Neovascularization Associated with Punctate Inner Choroidopathy: Combination of Intravitreal Anti-VEGF and Systemic Immunosuppressive Therapy.

PURPOSE: Choroidal neovascularization (CNV) associated with punctate inner choroidopathy (PIC) is a rare clinical entity, yet still a challenge for medical treatment. A case of a young myopic woman developing CNV secondary to unilateral PIC is presented. Clinical morphology, diagnostic procedure and follow-up are reported. CASE REPORT: A 29-year-old woman presented with multiple yellowish dots at the posterior pole. No other signs of inflammation could be seen. Angiography with fluorescein yielded hyperfluorescent signals in the affected areas with a diffuse leak, and SD-OCT showed a slightly elevated retinal pigment epithelial layer, consistent with the diagnosis of PIC. Additionally a classic CNV was observed. RESULTS: Anti-inflammatory therapy with local prednisolone acetate eye drops in combination with intravitreal injection of anti-vascular endothelial growth factor (VEGF, bevacizumab) yielded an increased best-corrected visual acuity. As CNV reappeared, systemic medication with prednisone and azathioprine in combination with two further intravitreal injections of anti-VEGF stabilized CNV and increased visual acuity again. CONCLUSION: Combined therapy of immunosuppression with intravitreal anti-VEGF injections can be considered as therapeutic strategy in the management of recurrent CNV associated with PIC.

Ocular diseases in metastatic cutaneous melanoma: review of 108 consecutive patients in two German tertiary centers.

PURPOSE: To analyze the incidence and spectrum of ocular disease in patients with metastatic cutaneous melanoma. METHODS: One hundred and eight consecutive patients with metastatic cutaneous melanoma were screened for ocular diseases using standardized eye examination, including measurement of visual acuity and intraocular pressure, slit-lamp examination, funduscopy in mydriasis, and spectral-domain optical coherence tomography (SDOCT) of the retina. Selected cases with atypical findings underwent electrophysiological studies. One patient was examined for hypercortisolism by a dexamethasone suppression test. RESULTS: Ocular diseases were found in 65 out of 108 patients (60 %) with metastatic cutaneous melanoma, significantly more often in older patients (p = 0.004). Cataract was present in 27 patients (25 %), pseudophakia in 22 patients (20 %), macular disease in 29 patients (28 %), diabetic retinopathy in ten patients (9 %), hypertensive retinal disease in 14 patients (13 %), retinal venous and arterial occlusive disease in three patients (3 %), optic neuropathy in four patients (4 %), and uveitis in one patient (1 %). Eight patients (8 %) had choroidal or iridal nevi, one patient (1 %) choroidal hemangioma, and one patient (1 %) choroidal metastasis. No patient had periocular neoplastic lesions. Paraneoplastic retinopathy manifesting as acute exudative polymorphous vitelliform maculopathy (AEPVM)-like disease was diagnosed in two patients (2 %) with multifocal central serous chorioretinopathy and development of vitelliform or fibrin-like subretinal deposits in one patient. CONCLUSIONS: Patients with metastatic cutaneous melanoma reveal ocular diseases with a spectrum similar to the normal population of this age range. Very rarely, uveal metastasis as well as paraneoplastic retinopathy can occur.

Topical Ranibizumab inhibits inflammatory corneal hem- and lymphangiogenesis.

PURPOSE: Ranibizumab (Lucentis(®) ) is a Fab-Fragment of a recombinant, humanized, monoclonal VEGF (anti-vascular endothelial growth factor) antibody. This study analyzed the ability of topical Ranibizumab to inhibit lymphangiogenesis in addition to hemangiogenesis after acute corneal inflammation in vivo. In addition, the effect of Ranibizumab on the proliferation of human lymphatic endothelial cells (LECs) and blood endothelial cells (BECs) in vitro was studied. METHODS: The inhibitory effect of Ranibizumab on LECs and BECs was studied in vitro using a proliferation enzyme-linked immunosorbent assay (ELISA) assay. To study the in vivo effects of Ranibizumab, the mouse model of suture induced inflammatory corneal neovascularization was used. Study mice received topical Ranibizumab as eye drops. After 1 week excised corneas were stained with LYVE-1 and CD31. Hemangiogenesis and lymphangiogenesis were analyzed morphometrically by using a semiautomatic method based on the image analyzing program Cell^F. RESULTS: An antiproliferative effect of Ranibizumab was seen in vitro on both human BECs and LECs with a significance of p < 0.0001 and p < 0.0004, respectively. In vivo experiments showed that topical application of Ranibizumab significantly inhibits both hemangiogenesis (p = 0.0026) and lymphangiogenesis (p = 0.0026) in the cornea. CONCLUSION: Ranibizumab is a potent inhibitor of inflammatory corneal hemangiogenesis and lymphangiogenesis in vivo with a direct inhibitory effect on both endothelial cell types in vitro. This study for the first time demonstrates an inhibitory effect of Ranibizumab on lymphatic vessels which could have a wider range of clinical applications.

Innervation pattern of the preocular human central retinal artery.

The central retinal artery (CRA) is the main vessel for inner retinal oxygen and nutrition supply. While the intraocular branches lack autonomic innervation, the innervation pattern of the extra-ocular part of this vessel along its course within the optic nerve is poorly investigated. This part however is essential for maintenance of retinal blood supply, in physiological and pathological conditions. Therefore, the aim of this study was the characterization of the autonomic innervation of the preocular CRA in humans with morphological methods. Meeting the Declaration of Helsinki, eyes of body or cornea donors were processed for single or double immunohistochemistry against tyrosine hydroxilase (TH), dopamine-ß-hydroxylase (DBH), choline acetyl-transferase (ChAT), vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (nNOS), calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), and cytochemistry for NADPH-diaphorase (NADPH-d). For documentation, light-, fluorescence-, and confocal laser-scanning microscopy were used. TH and DBH immunoreactive nerve fibres were detected in the CRA vessel wall, although a distinct perivascular plexus was missing. Further, nerve fibres immunoreactive for ChAT and VAChT were found, while CGRP, SP, and VIP were not detected. NADPH-d staining revealed scattered nerve fibres in the adventitia of the CRA and in close vicinity; however, nNOS-immunostaining could not confirm this finding. The CRA receives adrenergic and cholinergic innervations, indicating sympathetic and parasympathetic components, respectively. Remarkably, a peptidergic primary afferent innervation was missing. Since clinical results suggest an autoregulation of intraretinal vessels, further studies are needed to clarify the impact of CRA innervation for retinal perfusion.

Melatonin analogue agomelatine reduces rabbit's intraocular pressure in normotensive and hypertensive conditions.

In the search for new compounds to reduce intraocular pressure (IOP), with fewer side effects, we have found that agomelatine, a melatonin analogue, can reduce IOP being, therefore, interesting for the treatment of ocular hypertension and glaucoma. In normotensive conditions, agomelatine (10µl 100µM) reduced IOP by 20.8±1.4% (n=18) with a maximal effect 180min after the compound application and 68.8±5.7% (n=8) in a hypertensive condition. Concentration-response curve depicted a sigmoid behaviour presenting a pD2 value of 9.7±0.3 which was equivalent to an EC50 of 0.19nM. The effect of agomelatine was partially antagonized by 4PPDOT (MT2 antagonist receptor. 10µl 100µM) and prazosin (MT3 antagonist receptor. 10µl 100µM) (85.6±1.6% and 87.2±1.9%, N=18 respectively.) Agomelatine hypotensive effect in normotensive condition was comparable to latanoprost (40µl) and brimonidine (40µl) and it was no so effective as dorzolamide (40µl) or timolol (40µl). These results may suggest the use of this melatonin analogue for the treatment of those ocular conditions, which involve an abnormal raise of intraocular pressure.